FIG. 3.

CFTR-Δf508-derived hFSCs can differentiate into lung epithelium. (A) QPCR analysis showing that hFSCs derived from patients with the CFTR mutation Δf508 can differentiate into early lung endoderm and express early markers (NKX2.1, FOXP2, GATA6, and FOXA2) while they are negative for more mature markers (FOXJ1, SOX2, and SOX17) and thyroid markers (HHEX). (B) QPCR analysis showing that CFTR mutant lung progenitors can develop into mature airway epithelium and continue to express distal airway markers (NKX2.1, GATA6) as well as AECTII markers (SFTPB, SFTPC, and ABCA3) and AECTI markers (AQP5, P2X7, and PDPN). (C) Immunocytochemistry showing distal airway epithelium expresses NKX2.1, SFTPC, and CK18. (D) Confocal microscopy showing localization of Pro-SFTPC in the cytoplasm of the distal airway epithelium. (E) Trace of chloride influx and efflux in mature airway epithelium from CFTR-human-induced pluripotent stem cell showing the temperature sensitivity of CFTR. Chloride influx and efflux was measured in mature airway epithelium expressing the wild-type CFTR (green trace) cultured at 37°C, the Δf508 mutation (red trace) cultured at 37°C, and the Δf508 mutation (blue trace) cultured at 28°C. Addition of Cl⁻ or NO₃⁻ indicated with solid arrowheads. White bars=100 μM. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001. FL, human fetal lung control; AL, adult lung control; C, undifferentiated hESC control; Δf508, airway epithelium from cystic fibrosis patients cultured for 25 days.